Non-aqueous taxane formulations and methods of using the same

ABSTRACT

Non-aqueous, ethanol-free taxane formulations are provided. Formulations of embodiments of the invention include a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent. Also provided are methods of using the formulations, as well as kits that include the formulations. Non-aqueous, ethanol-free docetaxel formulations are provided. Formulations of embodiments of the invention include docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid which is soluble in the non-aqueous solvent and is substantially free of any conjugate base. Also provided are methods of using the formulations, as well as kits that include the formulations.

INTRODUCTION

This application is a Continuation of U.S. application Ser. No. 14/564,384, filed Dec. 9, 2014, now U.S. Pat. No. 9,308,195, which is a Continuation of U.S. application Ser. No. 14/041,675, filed Sep. 30, 2013, now U.S. Pat. No. 8,940,786, which claims priority from U.S. Provisional Applications 61/708,595 and 61/708,586, both filed Oct. 1, 2012, and each incorporated herein by reference in its entirety.

Taxanes constitute a family of naturally occurring diterpene compounds including paclitaxel. Paclitaxel, originally isolated from the bark of the Pacific Yew tree (Taxus brevifolia), and its semi-synthetic analogue, docetaxel, are two examples of taxane compounds. Taxanes are active agents that block cell growth by stopping mitosis via microtubule interference.

Taxanes can be used effectively to treat a variety of cancers and have been reported to have therapeutic effects in treating certain inflammatory diseases. Paclitaxel, for example, has been found to have activity against ovarian and breast cancers, as well as against malignant melanoma, colon cancer, leukemias and lung cancer (see, e.g., Borman, Chemical & Engineering News, Sep. 2, 1991, pp. 11-18; The Pharmacological Basis of Therapeutics (Goodman Gilman et al., eds.), Pergamon Press, New York (1990), p. 1239: Suffness, Antitumor Alkaloids, in: “The Alkaloids, Vol. XXV,” Academic Press, Inc. (1985), Chapter 1, pp. 6-18: Rizzo et al., J. Pharm. & Biomed. Anal. 8(2):159-164 (1990); and Biotechnology 9:933-938 (October, 1991).

Formulation of taxanes in therapeutically useful carriers, so as to enable the taxanes to be administered to animals, is made difficult by the nature of the taxane molecule, which can be poorly soluble in both aqueous and lipid carriers.

Docetaxel is an antineoplastic agent belonging to the taxoid family. It is prepared by semisynthesis beginning with a precursor extracted from the renewable needle biomass of yew plants. The chemical name for docetaxel is (2R,3S)—N-carboxy-3-phenylisoserine,Ntert-butyl ester, 13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one 4-acetate 2-benzoate. Decetaxcel has the following structural formula:

Docetaxel can be used effectively to treat a variety of cancers and has been reported to have a therapeutic effect in treating certain inflammatory diseases. For example, docetaxel has been found to have activity against breast cancer, non-small cell lung cancer, prostate cancer, gastric adenocarcinoma and head and neck cancer.

However, docetaxel is highly lipophilic and practically insoluble in water. This makes the formulation of docetaxel in therapeutically useful carriers a challenge.

SUMMARY

Non-aqueous, ethanol-free taxane liquid nanodispersion formulations are provided. The nanodispersion formulations include a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent. Also provided are methods of using the nanodispersion formulations, as well as kits that include the nanodispersion formulations.

Non-aqueous, ethanol-free taxane liquid nanodispersion formulations are provided. The nanodispersion formulations include a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent. Also provided are methods of using the nanodispersion formulations, as well as kits that include the nanodispersion formulations. Additionally, provided herein is a non-aqueous, ethanol-free docetaxel liquid nanodispersion formulation comprising docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base. Also provided are methods of using the nanodispersion formulations, as well as kits that include the nanodispersion formulations.

DETAILED DESCRIPTION Taxane Nanodispersion Formulations

Non-aqueous, ethanol-free taxane liquid nanodispersion formulations are provided. The nanodispersion formulations include a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent. Also provided are methods of using the nanodispersion formulations, as well as kits that include the nanodispersion formulations.

Before the present invention is described in greater detail, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

Certain ranges are presented herein with numerical values being preceded by the term “about”. The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative illustrative methods and materials are now described.

All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

It is noted that, as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely”, “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

In the following sections, the taxane nanodispersion formulations and taxane diluted solutions prepared therefrom, as well as methods using the same, are described in greater detail, as well as methods for preparing the nanodispersion formulations and diluted solutions, as well as kits that may include the formulations.

Aspects of the invention include taxane nanodispersion formulations. In some instances, the nanodispersion formulations are non-aqueous liquid, ethanol-free compositions that, upon combination with an aqueous medium, produce a taxane diluted solution. The non-aqueous, ethanol-free taxane liquid nanodispersion formulations of embodiments of the invention comprise a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent. In certain embodiments, the non-aqueous, ethanol-free taxane liquid nanodispersion formulations of embodiments of the invention consist essentially of a taxane, an oil, a non-ionic surfactant, a non-aqueous solvent, and an organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent.

As used herein, the term “nanodispersion” refers a substantially clear (i.e., transparent) solution comprising nanosized particles. In some instances, the particle size ranges from about 3 to about 70 nm, from about 5 to about 50 nm, from about 7 to about 30 nm, or from about 8 to about 15 nm.

Taxanes of interest are diterpene compounds. In some instances, taxanes are compounds described by the formula:

Of interest are anhydrous taxanes as well as hydrates thereof, e.g., mono, di, tri, tetra and penta hydrates, etc. In certain embodiments, the taxane is paclitaxel or docetaxel, including anhydrous or a hydrate thereof, e.g., anyhydrous docetaxel, docetaxel trihydrate, paclitaxel trihydrate, etc., may be employed in the formulations. Taxanes of interest also include, but are not limited to: 7-epitaxol, 7-acetyl taxol, 10-desacetyl-taxol, 10-desacetyl-7-epitaxol, 7-xylosyltaxol, 10-desacetyl-7-glutaryltaxol, 7-N,N-dimethylglycyltaxol, 7-L-alanyltaxol, SB-T-1011, etc. The taxane may be present as a free base or salt.

Taxane nanodispersion formulations include an effective amount of a taxane. By effective amount is meant a dosage sufficient to provide the desired result, e.g., inhibition of cellular proliferation. The effective amount of taxane may vary depending on the particular taxane employed, and in certain embodiments ranges from 0.05 to 5% by weight, such as 0.5 to 5% by weight and including 0.3 to 3% by weight. In certain embodiments, the nanodispersion formulations include an effective amount of paclitaxel or paclitaxel trihydrate. In certain embodiments, paclitaxel or paclitaxel trihydrate is present in the nanodispersion formulation in an amount ranging from 0.05 to 5.0% w/w, such as 0.5 to 5.0% w/w, and including 0.3 to 3.0% w/w, where in some instances the amount ranges from 0.3 to 5.0% w/w, such as 0.3 to 3.0% w/w, e.g., 0.4 to 2.5% w/w, e.g., 0.5 to 2.0% w/w, and including 1.0 to 1.5% w/w. In certain embodiments, the nanodispersion formulations include an effective amount of docetaxel or docetaxel trihydrate. In certain embodiments, anhydrous docetaxel or docetaxel trihydrate is present in the nanodispersion formulation in an amount ranging from about 0.1 to about 5 wt %, or from about 0.5 to about 5 wt %, or from about 0.5 to about 3 wt %, or from about 1 to about 3 wt %, or is about 2 wt %. The concentration of the taxane in the formulations may vary, and is in some embodiments, about 30 mg/ml or less, about 25 mg/ml or less, about 20 mg/ml or less, about 10 mg/ml or less, about 1 mg/ml or less, or ranges from about 0.05 to about 20 mg/ml, from about 0.5 to about 20 mg/ml, from about 1 to about 20 mg/ml, or from about 5 to about 20 mg/ml.

As used herein, the term “non-aqueous” is intended to refer to formulations having a water content about 3% by weight or less. The water content may come from any one or more of the components in the formulation, such as the organic acid component or from the water associated with docetaxel trihydrate, when used. In some embodiments, the formulations comprise less than about 3% water by weight, or less than about 2.5% water by weight, or less than about 2% water by weight, or less than about 1.5% water by weight, or less than about 1% water by weight, or less than about 0.5% water by weight, or less than about 0.1% water by weight, or less than about 0.01% water by weight, or than about 0.005% water by weight.

Also present in the nanodispersion formulations is an oil component made up of one or more oils. Without wishing to be bound by any one theory, it is contemplated that the inclusion of the oil in the nanodispersion formulation helps stabilize the diluted solution. Oils of interest are physiologically acceptable and include, but are not limited to: simple lipids, derived lipids, complex lipids that are derived from vegetable oil and fat, animal oil and fat, or mixtures thereof, where the oils may be naturally occurring or synthetic.

In certain embodiments, the oil includes, but is not limited to vegetable oil such as soybean oil, olive oil, sesame oil, castor oil, corn oil, peanut oil, safflower oil, grape seed oil and eucalyptus oil, medium-chain fatty acid esters, low-chain fatty acid esters, triglycerides, and the like. Animal oils and fat of interest include, but are not limited to, cod-liver oil, seal oil, sardine oil, docosahexiaenoic acid, and eicosapentaenoic acid. One or a combination of more than one of these types of oils can be used. For example, some embodiments of the subject formulations include soybean oil, olive oil, sesame oil, or combinations thereof. Other embodiments include soybean oil, olive oil, or combinations thereof. Highly refined oils and fats are employed in certain embodiments. In some embodiments, the oil is soybean oil. In some embodiments, the oil is a medium chain triglyceride (i.e., a medium chain fatty acid ester). In some embodiments, the oil is not a medium chain triglyceride (i.e., a medium chain fatty acid ester).

Oils to be used in the formulations disclosed herein may also include tocopherols. Tocopherols are a family of natural and synthetic compounds, also known by the generic names tocols or Vitamin E. α-Tocopherol is the most abundant and active form of this class of compounds and it has the following chemical structure:

Other members of this class include α-, β-, γ-, and δ-tocotrienols, and α-tocopherol derivatives such as tocopherol acetate, phosphate, succinate, nitotinate and linoleate. Any convenient tocopherol may be present, as desired, including the specific tocopherols listed above.

Oils of interest also include polyol esters of medium chain fatty acids. The term “polyol esters of medium chain fatty acids” is intended to include esters and mixed esters of glycerol, propylene glycol or other open chain polyols such as polyethylene glycol, reacted with medium chain fatty acids, e.g., where the acid has a chain length between 6 and 12 carbon atoms. In some instances, the polyol esters of medium chain fatty acids are triglycerides or diglycerides of the C₈-C₁₀ fatty acids, e.g., as may be commercially available from the fractionation of coconut oil. Commercially available products of this description are sold under the trade names “NEOBEE®”, “ODO®”, “Panacet®”, “Miglyol®” and “Captex® 300” and have a majority of C₈ fatty acid (caprylic) triglyceride and C₁₀ fatty acid (capric) triglyceride with minor levels of C₆ and C₁₄ fatty acid triglycerides. In some embodiments, the oil is NEOBEE®.

In certain embodiments, the oil is selected from the group consisting of synthetic oils, vegetable oils, tocopherols and combinations thereof. In other embodiments, the oil is selected from the group consisting of soybean oil, olive oil, sesame oil, corn oil, a medium chain triglyceride, a tocopherol or derivative thereof and combinations thereof.

In certain embodiments, the oil is about 20 wt % or less of the formulation. In some instances, the amount of oil in the nanodispersion formulation ranges from about 0.5 to about 20 wt %, or from about 1 to about 20 wt %, or from about 1 to about 10 wt %. In certain embodiments, the oil is present in an amount ranging from about 1 to about 5 wt %. In other embodiments, the oil is present in an amount ranging from about 1 to about 3 wt %, or about 2 wt %. In certain embodiments, the amount of oil in the nanodispersion formulation is calculated based on the amount of docetaxel in the formulation. For example, in some embodiments, when the amount of oil greatly exceeds the amount of docetaxel (w/w), the particle size can become larger, which typically results in the formulation becoming cloudy. Accordingly, in some embodiments, the amount of oil is less than about three times the amount of taxane (w/w), or equal to or less than about 2.5 times the amount of taxane (w/w), or equal to or less than about 2 times the amount of taxane (w/w), or equal to or less than about 1.5 times the amount of taxane (w/w), or is about equal to the amount of taxane (w/w), or is about half of the amount of taxane (w/w).

Also present in the nanodispersion formulations is a non-ionic surfactant, which may include one or more non-ionic surfactants. Surfactants of interest include any type of non-ionic surfactant that can be used for pharmaceutical formulations. Non-ionic surfactants of interest include, but are not limited to, polyoxyalkylene copolymer, and sorbitan fatty acid esters. In some embodiments, the sorbitan fatty acid ester is a polyoxyethylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan tristearate (Tween® 65); polyoxyethylene sorbitan trioleate (Tween® 85); polyethylene glycol 400 monostearate; polysorbate 60; (Tween® 60); polyoxyethylene monostearate (Myrj® 49); polysorbate 80 (Tween® 80); polysorbate 40 (Tween® 40); and polysorbate 20 (Tween 20)) or sorbitan fatty acid esters (e.g., sorbitan trioleate (Span® 85); sorbitan tristearate (Span® 65); sorbitan sesquioleate (Arlacel® 83); glyceryl monostearate; sorbitan monooleate (Span® 80); sorbitan monostearate (Span® 60); sorbitan monopalmitate (Span® 40); and sorbitan monolaurate (Span® 20)). In some embodiments, the non-ionic surfactant is polysorbate 80. In some embodiments, the polysorbate 80 is refined grade.

The amount of non-ionic surfactant in the nanodispersion formulation may vary. In some instances, the amount of non-ionic surfactant in the nanodispersion formulation is 40 wt % or more. In some instances, the non-ionic surfactant is present in an amount ranging from about 40 to about 75 wt %, or from about 50 to about 65 wt %, or from about 50 to about 60 wt %, or from about 50 to about 57 wt %, or from about 57 to about 65 wt %. The combination ratio by weight of the oil and the surfactant in the subject nanodispersion formulations may vary, and is in some instances about 1/100, or 1/50, or 1/40, or 1/30, or 1/20, or 1/10, or 1/8, or 1/6, or 1/4, or 1/2, or 1/1.

The nanodispersion formulations of the invention further include a non-aqueous solvent, which may include one or more non-aqueous solvents. Non-aqueous solvents of interest include, but are not limited to, propylene glycol, polypropylene glycol, polyethylene glycol (such as PEG 300, PEG 400, PEG 600, PEG 800, PEG 1000, etc., where in certain embodiments polyethylene glycols, when employed, have an average molecular weight of 1000 or less), glycerin, triacetin, dimethyl isosorbide, glycofurol, propylene carbonate, dimethyl acetamide or a mixture thereof. In some embodiments, the non-aqueous solvent is polyethylene glycol. In some embodiments, the non-aqueous solvent is polyethylene glycol 400. In some embodiments, the polyethylene glycol has a melting point of less than 0° C. In some embodiments, the non-aqueous solvent is polyethylene glycol 300.

The non-aqueous solvent may be present in varying amounts, and in some instances is present in an amount ranging from about 20 to about 60 wt %, or from about 35 to about 45 wt %, including amounts ranging from about 35 to about 40 wt %, or from about 40 to about 45 wt %.

In the present formulations, the amount of non-ionic surfactant (w/w) should be about equal to or greater than the amount of non-aqueous solvent (w/w) in the formulation, or greater than about 95% of the amount of non-aqueous solvent (w/w), or in other words, the ratio by weight of the non-ionic surfactant and the non-aqueous solvent is 0.95 or more. As shown in the Examples, when used in such amounts, the stability of the formulation is enhanced.

In the present nanodispersions, the combination ratio by weight of the oil and the non-aqueous solvent in the nanodispersion formulations may vary, and in some instances is 1/100, or 1/50, or 1/40, or 1/30, or 1/20, or 1/10, or 1/7, or 1/5, or 1/3, or 1/1.

The nanodispersion formulation further comprises an organic acid component. An organic acid component may include an organic acid and/or an organic acid buffer (i.e., an organic acid and its conjugate base (or salt thereof)). Therefore, in some instances, the organic acid buffer comprises an organic acid and a salt of its conjugate base. Organic acids of interest upon which the organic acid component may be based include those which are soluble in the non-aqueous solvent (in the amount used). Specific examples include lactic acid, succinic acid, malic acid, tartaric acid and acetic acid.

In some instances, the organic acid component is a lactic acid/sodium lactate component, such that the component includes both lactic acid and sodium lactate. As organic acid can be hydroscopic and may contain a small or trace amount of water, the lactic acid and/or sodium lactate can be commercially available or substantially dehydrated prior to use. In some instances, the lactic acid/sodium lactate component is present in an amount ranging from about 0.3 to 3 wt %, or from about 0.3 to about 2 wt %, or from about 0.3 to about 1.5 wt %, or from about 0.5 to about 1.5 wt %, or from about 0.8 to about 1.2 wt %, or from about 0.8 to about 1 wt %, or about 1 wt %.

In other instances, the organic acid component, e.g., lactic acid or acetic acid, does not contain any significant amount of its conjugate base (or salt thereof). In addition, in some embodiments, the nanodispersion formulations disclosed herein do not contain a buffer or a mineral acid. In some embodiments, the organic acid component is lactic acid. The lactic acid can be D- or L-lactic acid, or a mixture thereof. The organic acid may be present in varying amounts, and in some instances is present in an amount ranging from about 0.3 to about 3 wt %, or from about 0.3 to about 1 wt %, or from about 0.3 to about 0.6 wt %, or from about 0.4 to about 0.5 wt %, or from about 0.5 to about 1 wt %, or from about 0.7 to about 1 wt %, or about 0.8 wt %.

In some embodiments, the present disclosure provides for a non-aqueous, ethanol-free docetaxel liquid nanodispersion formulation comprising: docetaxel, soybean oil in an amount ranging from about 1 to about 5 wt %, polysorbate 80 in an amount ranging from about 50 to about 60 wt %, polyethylene glycol in an amount ranging from about 35 to about 45 wt %, and lactic acid or lactic acid buffer in an amount ranging from 0.3 to 1 wt %, wherein the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent.

In another embodiment, the present disclosure provides for a non-aqueous, ethanol-free docetaxel liquid nanodispersion formulation comprising: docetaxel, a medium chain triglyceride in an amount ranging from about 1 to about 5 wt %, polysorbate 80 in an amount ranging from about 50 to about 60 wt %, polyethylene glycol in an amount ranging from about 35 to about 45 wt %, and lactic acid or lactic acid buffer in an amount ranging from 0.3 to 1 wt %, wherein the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent.

The nanodispersion formulation as described above typically has a pH of less than about 4, or less than about 3.5, or less than about 3.4, or less than about 3.3, or less than about 3.25, or less than about 3.2, or less than about 3.1, or from about 3.0 to about 3.1.

Methods of Preparing Taxane Nanodispersion Formulations

Nanodispersion formulations may be prepared according to any convenient protocol. As such, the components of the desired nanodispersion may be combined under conditions sufficient to produce the desired nanodispersion. Accordingly, an amount of taxane, oil, non-ionic surfactant, non-aqueous solvent, and organic acid component, wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent, may be combined under conditions sufficient to produce a nanodispersion. The components may be combined in any convenient order. The components may be combined at any convenient temperature, e.g., room temperature or elevated temperatures, such as temperatures ranging from 30 to 95° C., or 50 to 60° C., or 70 to 80° C. Certain of the components may be combined with each other, and then combined together, or all of the components may be combined at substantially the same time. Combination may include various manners of agitation or mixing, e.g., stirring, sonication, etc., in order to produce the nanodispersion. Depending on the particular preparation method, an aqueous solvent, e.g., water, etc. may or may not be employed during preparation of the nanodispersion compositions.

In one embodiment, a nanodispersion is prepared without an aqueous solvent. In these embodiments, the desired amounts of components of the nanodispersion, e.g., taxane, oil, non-ionic surfactant, non-aqueous solvent, and organic acid component are combined. Combination may include various manners of agitation, e.g., stirring, sonication, etc., in order to produce the nanodispersion. Where desired, heat may be employed to facilitate mixing, although, in some embodiments, the combining is performed at ambient temperature. In certain embodiments, the nanodispersion is clear. By clear is meant that the nanodispersion is a translucent, if not transparent liquid, i.e., the liquid is pellucid.

In some instances, the nanodispersion preparation protocol includes use of an aqueous solvent, e.g., pure water. In these instances, an aqueous formulation is prepared, which includes taxane, oil, non-ionic surfactant, non-aqueous solvent, organic acid component and an aqueous solvent, e.g., water, etc. In certain embodiments, the aqueous formulation composition is clear. By clear is meant that the nanodispersion is a translucent, if not transparent liquid, i.e., the liquid is pellucid. As such, the initial preparation is not cloudy. In some instances, the particle size of the initial composition ranges from about 3 to about 70 nm, such as about 5 to about 50 nm and including about 7 to about 30 nm, such as about 8 to about 15 nm. Of interest in certain embodiments are initial compositions that are clear (e.g., as described above) and have a particle size of about 70 nm or less, such as about 50 nm or less, including about 30 nm or less, including about 25 nm or less, about 20 nm or less and about 15 nm or less. In these embodiments, as a final step, water may be removed from the composition to produce a final, non-aqueous nanodispersion. Removal of water may be accomplished using any convenient protocol, e.g., via a combination of pressure and/or temperature modulation, such as heating.

The preparation methods can be carried out at certain temperature such as temperatures ranging from about 30° C. to about 95° C., about 50° C. to about 60° C., about 60° C. to about 70° C., or about 70° C. to about 80° C. Specific examples are provided in the Experimental section, below.

In some embodiments, the method for making the ethanol-free taxane nanodispersion formulation as disclosed herein does not comprise the addition of water and/or ethanol, followed by removal thereof. Accordingly, since the non-aqueous, ethanol-free taxane nanodispersion formulation has not been contacted with either water or ethanol during the formulation process, it is substantially free of water (i.e, less than about 3%) and free of ethanol. In some embodiments, the formulation is essentially free of water (i.e, less than about 1%). In addition, in certain embodiments, the formulations do not have any detectable amount of ethanol. This results in a formulation which is stable and can be suitable for subjects having an adverse reaction to ethanol. In some embodiments, the formulations do not contain activated carbon.

Where desired, an amount of the nanodispersion may be loaded into an individual dosage container, e.g., vial, which holds the nanodispersion and keeps it sterile during shipping, storage, and handling. Before or during the loading stage, the nanodispersion can be passed through a sub-micron sterilizing filter, e.g., a 0.2μ hydrophilic filter) which has a sufficiently small pore size to remove any bacteria or viruses. The sterilizing filters of interest include, but are not limited to hydrophilic filters. In some embodiments, the filter could be a CA (Cellulose Acetate) membrane filter, PTFE (Polytetrafluoroethylene) membrane filter, PVDF (Polyvinylidene fluoride or polyvinylidene difluoride) membrane filter or PES (Polyethersulfone) membrane filter.

As used herein, the term “vial” refers to any container that is used to hold the nanodispersion formulation. Many pharmaceutical vials are made of clear glass, which allows several advantages, including visual inspection of the enclosed drug (to ensure that it is still in a clean, non-caramelized, non-collapsed form, when it is ready for use) and of the container itself (to ensure that it does not have a hairline crack in one of the walls, which could jeopardize or destroy sterility of the enclosed drug). Various types of pharmaceutical vials are known. Single-chamber vials can be sealed with rubber or plastic plugs that will allow a hypodermic needle to be pushed through the rubber seal. Examples include any hydrolytically stable glass, such as a borosilicate Type I or soda-lime-silica Type II glass vial, having a suitable stopper (e.g., Teflon coating). Alternately, a single-chamber vial can be made of a brittle and easily breakable material, inside a sealed bag that can contain an aqueous solution (such as physiological saline or a dextrose solution, in an intravenous infusion bag); if this type of vial is broken, it will release its contents into the still-sealed bag, for mixing. In yet other embodiments, two-chamber vials or analogous structures, e.g., as described in Published United States Application Publication No. 2003/0099674 and U.S. Pat. No. 4,781,354 may be employed. Other methods for preparing the taxane formulations disclosed herein can be found in Published United States Application Publication No. 2011/0269829, the entirety of which is incorporated herein by reference.

Where desired, the nanodispersion formulation may be stored for a period of time prior to combination with the aqueous medium. This storage time of the nanodispersion may vary, where storage times may be 1 year or more, such as 2 years or more, including 3 years or more. While the storage conditions may vary, in certain instances the storage conditions are characterized by a temperature ranging from about 5 to 60° C., e.g., 5° C., such as about 8 to 40° C., e.g., about 25° C. The activity of the taxane active agent is substantially preserved during the storage period, such that the nanodispersion formulations are storage stable. As such, the activity of the taxane active agent in the infusion solution following storage is substantially the same as that in the nanodispersion prior to being dried, where the magnitude of any difference in activity between the nanodispersion and diluted solution may be about 15% or less, such as about 10% or less, including about 5% or less, e.g., as can be measured by HPLC.

As shown in the Examples, the nanodispersion formulation according the present disclosure is stable for at least 6 months (see Examples 3 and 6) at 40° C. The stability of the present nanodispersion formulations can be determined by methods known in the art, such as by measuring the recovery rate for the taxane (e.g., docetaxel) peak by HPLC. In some embodiments, the nanodispersion formulation exhibits a taxane (e.g., docetaxel) recovery rate of more than about 95% after 6 months at about 40° C., or about 96% or more, or more than about 97% or more, or more than about 98% or more, or about 99% or more. The recovery rate is calculated based on the measured amount of docetaxel in the formulation, which is not necessarily the amount of docetaxel added to the formulation (thus eliminating any potential error from an impurity in the docetaxel).

Taxane Diluted Solutions and Methods of Use

Following preparation of the nanodispersion formulation, e.g., as described above, at the time of desired administration to a subject, a dosage amount of the nanodispersion may be combined with an aqueous medium to prepare an diluted solution that is suitable for use. The dosage amount of the nanodispersion formulation may be combined with any suitable aqueous medium, where aqueous mediums of interest include, but are not limited to: deionized water, USP water for injection (WFI), salines, transfusion solutions, physiological solutions, etc. In some embodiments, the aqueous medium comprises an aqueous 0.9% sodium chloride solution or an aqueous 5% dextrose or glucose solution. The liquids to nanodispersion (high viscous liquid) ratio employed during preparation of the diluted solution may vary, and in certain embodiments ranges from about 0.5 to about 300, such as about 1 to about 100, about 2 to about 50 or about 2 to about 20, and including about 2 to about 10. In some instances, the dosage amount of nanodispersion formulation that is combined with the aqueous medium ranges from about 100 to about 1200 g, such as about 300 to about 600 g and the amount of aqueous medium that is combined with the dosage amount ranges from about 100 to about 1200 ml, such as about 250 to about 600 ml.

The diluted solution prepared from the nanodispersion formulations are liquid preparations that are a suspension of small particles (i.e., globules) of one liquid in a second liquid with which the first liquid will not mix. The water present in the taxane diluted solutions may be any convenient water, including deionized water, USP water for injection (WFI), etc.

The combination protocol may vary, where agitation may be employed, e.g., by stirring, by kneading a bag that includes both the nanodispersion and the aqueous medium, etc.

The diluted solutions include a taxane, an oil component, a non-ionic surfactant component, a non-aqueous solvent component and an aqueous medium. In certain embodiments, the diluted solutions are clear. By clear is meant that the diluted solution is a translucent, if not transparent liquid, i.e., the liquid is pellucid. As such, the diluted solution is not cloudy, e.g., as a suspension may appear. Further details regarding the diluted solutions that may be prepared from the taxane nanodispersion precursors are provided below. In some instances, the particle size of the final diluted solution ranges from about 3 to about 70 nm, such as about 5 to about 50 nm and including about 7 to about 30 nm, such as about 8 to about 15 nm. Of interest in certain embodiments are diluted solutions that are clear (e.g., as described above) and have a particle size of about 70 nm or less, such as about 50 nm or less, including about 30 nm or less, including about 25 nm or less, about 20 nm or less and about 15 nm or less. In some instances, any difference in particle size between the nanodispersion and diluted solutions is minimal, such that the particle sizes in the nanodispersion and diluted solutions are substantially the same. In some instances, any difference in particle size between the nanodispersion and diluted solutions is about 30 nm or less, such as about 20 nm or less, about 15 nm or less, about 10 nm or less, or about 5 nm or less. Without wishing to be bound by any one theory, it is contemplated that the inclusion of the oil in the nanodispersion formulations helps stabilize the diluted solution.

The diluted solutions have a physiologically acceptable pH. In certain embodiments, the pH of the diluted solutions ranges from about 2.5 to about 8, such as from about 3 to about 7, including from about 3.5 to about 6. The diluted solutions are substantially clear (i.e., transparent) formulations. The concentration of the taxane in the diluted solutions may vary, ranging in some embodiments from about 0.05 to about 10 mg/ml, such as about 0.2 to about 3 mg/ml.

Methods of using the diluted solutions include administering an effective amount of the diluted solutions to a subject in order to treat the subject for a target condition of interest. By “treating” or “treatment” is meant at least a suppression or an amelioration of the symptoms associated with the condition afflicting the subject, where suppression and amelioration are used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g., symptom, associated with the condition being treated, such as pain. As such, treatment also includes situations where the condition is completely inhibited, e.g., prevented from happening, or stopped, e.g., terminated, such that the subject no longer experiences the condition. As such, treatment includes both preventing and managing a condition.

In practicing the methods, the diluted solutions disclosed herein can be parenterally administered to a subject. By “parenteral administration” is meant administration by a protocol that delivers a quantity of the diluted solutions to the subject, e.g., a patient suffering from a cellular proliferative disease, by a route other than the digestive tract. Examples of parenteral administration include, but are not limited to, intramuscular injection, intravenous injection, and the like. In certain embodiments, parenteral administration is by injection using an injection delivery device.

The amount of diluted solution that is administered to the subject may vary depending on a number of factors, such as patient specifics, nature of condition, nature of taxane active agent, etc. In certain embodiments, the volume of diluted solution that is administered to a subject may range from about 100 to about 1000 ml, such as about 200 to about 600 ml. The time period over which this volume is administered may vary, ranging from about 0.5 to about 6 hr, such as from about 1 to about 3 hours. Dosages administered to a subject during a given procedure may also vary, ranging in some instances from about 20 to about 500 mg/m², such as from about 50 to about 300 mg/m².

Accordingly, provided herein are methods of administering an non-aqueous, ethanol-free taxane liquid nanodispersion formulation to a subject, the method comprising: (a) combining the non-aqueous, ethanol-free taxane liquid nanodispersion formulation according to the present disclosure with an aqueous medium to provide a diluted solution; and (b) administering the diluted solution to the subject.

In some embodiments, the method of using the nanodispersion formulation disclosed herein comprises the steps of: (a) aseptically withdrawing the desired amount of the nanodispersion formulation (such as a formulation comprising about 20 mg taxane/mL) with a calibrated syringe, (b) injecting said formulation into a 250 mL infusion bag or bottle of either 0.9% sodium chloride solution or 5% dextrose solution to provide a diluted solution having a final taxane concentration of from about 0.3 mg/mL to about 0.74 mg/mL, and (c) administering said diluted solution to a patient. If a dose greater than 200 mg of taxane is required, one may use a larger volume of the infusion vehicle so that a concentration of 0.74 mg/mL taxane is not exceeded.

In certain embodiments, the subject methods include a diagnostic step. Individuals may be diagnosed as being in need of the subject methods using any convenient protocol. In addition, individuals may be known to be in need of the subject methods, e.g., they are suffering from a target disease condition (e.g., cellular proliferative disease, prior to practicing the subject methods. Diagnosis or assessment of target condition can be performed using any convenient diagnostic protocol.

Methods of the invention may further include assessing the efficacy of the treatment protocol that includes administration of the taxane diluted solution. Assessing the efficacy of treatment may be performed using any convenient protocol.

Taxane diluted solutions of the invention may be administered to a variety of different types of subjects. Subjects of interest include, but are not limited to: mammals, both human and non-human, including the orders carnivore (e.g., dogs and cats), rodentia (e.g., mice, guinea pigs, and rats), lagomorpha (e.g. rabbits) and primates (e.g., humans, chimpanzees, and monkeys). In certain embodiments, the subjects, e.g., patients, are humans.

In determining whether to administer the diluted solution to a particular given subject, care will be taken to ensure that the formulation is not contraindicated for that subject. As such, symptoms of the subject may be assessed to ensure that administration of the diluted solution will not have adverse effects that outweigh any benefit that the diluted solution may provide.

Utility

The subject formulations, diluted solutions and/or methods find use in a variety of applications, including the treatment of subjects suffering from cellular proliferative disease conditions. Cellular proliferative diseases that may be treated with formulations of the invention include, but are not limited to: carcinomas, myelomas, neuroblastomas, or sarcomas, of the brain, breast, lung, colon, prostate or ovaries, as well as leukemias or lymphomas. Specific disease conditions of interest include, but are not limited to, human ovarian cancer, breast cancer, malignant lymphoma, lung cancer, melanoma, and Kaposi's sarcoma.

Kits

Also provided are kits that find use in practicing the subject methods, as described above. For example, kits for practicing the subject methods may include a quantity of the nanodispersion formulation, present in unit dosages, e.g., vials, or a multi-dosage format. As such, in certain embodiments, the kits may include one or more unit dosages (e.g., vials) of the nanodispersion formulation. The term “unit dosage”, as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of the subject nanodispersion formulation calculated in an amount sufficient to produce the desired effect. The amount of the unit dosage of the subject formulation depends on various factors, such as the particular active agent employed, the effect to be achieved, and the pharmacodynamics associated with the active agent in the subject. In yet other embodiments, the kits may include a single multi-dosage amount of the formulation.

In certain embodiments, the kits may further include an amount of an aqueous medium suitable for use in production of the diluted solution. The aqueous medium may be any convenient aqueous medium, such as described above, present in any suitable container, e.g., an IV bag.

In some embodiments, the kits may include a syringe which is suitable to prepare the docetaxel diluted solution. A syringe with graduations is preferred to measure a certain amount of the docetaxel nanodispersion.

In addition to the above components, the subject kits may further include instructions for practicing the subject methods. These instructions may be present in the subject kits in a variety of forms, one or more of which may be present in the kit. One form in which these instructions may be present is as printed information on a suitable medium or substrate, e.g., one or more pieces of paper on which the information is printed, in the packaging of the kit, in a package insert, etc. The instructions may be present on a computer readable medium, e.g., diskette, CD, DVD, etc., on which the information has been recorded. The instructions may be present on a website, which may be used via the internet to access the information at a removed site. Other convenient means are possible and may be included in the kits.

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

Docetaxel Nanodispersion Formulations

Aspects of the invention include docetaxel nanodispersion formulations. In some instances, the nanodispersion formulations are non-aqueous, ethanol-free docetaxel liquid compositions that, upon combination with an aqueous medium, produce a docetaxel diluted solution. The non-aqueous liquid nanodispersion formulations of embodiments of the invention comprise docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base. In certain embodiments, the non-aqueous liquid nanodispersion formulations of embodiments of the invention consists essentially of docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base.

As used herein, the term “nanodispersion” refers a substantially clear (i.e., transparent) solution comprising nanosized particles. In some instances, the particle size ranges from about 3 to about 70 nm, from about 5 to about 50 nm, from about 7 to about 30 nm, or from about 8 to about 15 nm.

Anhydrous docetaxel as well as hydrates thereof, e.g., mono, di, tri, tetra and penta hydrates, etc, may be employed in the docetaxel nanodispersion formulations. In certain embodiments, the docetaxel is anhydrous docetaxel. In other embodiments, the docetaxel is docetaxel trihydrate.

Nanodispersion formulations include an effective amount of docetaxel. By effective amount is meant a dosage sufficient to provide the desired result, e.g., inhibition of cellular proliferation. The effective amount of docetaxel may range from about 0.1 to about 5 wt %, or from about 0.5 to about 5 wt % (weight percent), or from about 0.5 to about 3 wt %, or from about 1 to about 3 wt %, or is about 2 wt %. Therefore, in certain embodiments, anhydrous docetaxel or docetaxel trihydrate is present in the nanodispersion formulation in an amount ranging from about 0.1 to about 5 wt %, or from about 0.5 to about 5 wt %, or from about 0.5 to about 3 wt %, or from about 1 to about 3 wt %, or is about 2 wt %. The concentration of the taxane in the formulations may vary, and is in some embodiments, about 30 mg/ml or less, about 25 mg/ml or less, about 20 mg/ml or less, about 10 mg/ml or less, about 1 mg/ml or less, or ranges from about 0.05 to about 20 mg/ml, from about 0.5 to about 20 mg/ml, from about 1 to about 20 mg/ml, or from about 5 to about 20 mg/ml.

As used herein, the term “non-aqueous” is intended to refer to formulations having a water content about 3% by weight or less. The water content may come from any one or more of the components in the formulation, such as the organic acid component or from the water associated with docetaxel trihydrate, when used. In some embodiments, the formulations comprise less than about 3% water by weight, or less than about 2.5% water by weight, or less than about 2% water by weight, or less than about 1.5% water by weight, or less than about 1% water by weight, or less than about 0.5% water by weight, or less than about 0.1% water by weight, or less than about 0.01% water by weight, or than about 0.005% water by weight.

Also present in the nanodispersion formulations is an oil component made up of one or more oils. Without wishing to be bound by any one theory, it is contemplated that the inclusion of the oil in the nanodispersion formulations helps stabilize the diluted solution. Oils of interest are physiologically acceptable and include, but are not limited to: simple lipids, derived lipids, complex lipids that are derived from vegetable oil and fat, animal oil and fat, and mineral oil, or mixtures thereof, where the oils may be naturally occurring or synthetic.

In certain embodiments, the oil includes, but is not limited to vegetable oil such as soybean oil, olive oil, sesame oil, castor oil, corn oil, peanut oil, safflower oil, grape seed oil and eucalyptus oil, medium-chain fatty acid esters, low-chain fatty acid esters, triglycerides, and the like. Animal oils and fat of interest include, but are not limited to, cod-liver oil, seal oil, sardine oil, docosahexiaenoic acid, and eicosapentaenoic acid. One or a combination of more than one of these types of oils can be used. For example, some embodiments of the subject formulations include soybean oil, olive oil, sesame oil, or combinations thereof. Other embodiments include soybean oil, olive oil, or combinations thereof. Highly refined oils and fats can be employed in certain embodiments. In some embodiments, the oil is soybean oil.

Oils to be used in the formulations disclosed herein may also include polyol esters of medium chain fatty acids. The term “polyol esters of medium chain fatty acids” is intended to include esters and mixed esters of glycerol, propylene glycol or other open chain polyols such as polyethylene glycol, reacted with medium chain fatty acids, e.g., where the acid has a chain length between 6 and 12 carbon atoms. In some instances, the polyol esters of medium chain fatty acids are triglycerides or diglycerides of the C₈-C₁₀ fatty acids, e.g., as may be commercially available from the fractionation of coconut oil. Commercially available products of this description are sold under the trade names NEOBEE®, “ODO®” Panacet®, “Miglyol®” and “Captex® 300” and have a majority of C₈ fatty acid (caprylic) triglyceride and C₁₀ fatty acid (capric) triglyceride with minor levels of C₆ and C₁₄ fatty acid triglycerides. In some embodiments, the oil is NEOBEE®.

In certain embodiments, the oil is about 20 wt % or less of the formulation. In some instances, the amount of oil in the nanodispersion formulation ranges from about 0.5 to about 20 wt %, or from about 1 to about 20 wt %, or from about 1 to about 10 wt %. In certain embodiments, the oil is present in an amount ranging from about 1 to about 5 wt %. In other embodiments, the oil is present in an amount ranging from about 1 to about 3 wt %, or about 2 wt %. In certain embodiments, the amount of oil in the nanodispersion formulation is calculated based on the amount of docetaxel in the formulation. For example, in some embodiments, when the amount of oil greatly exceeds the amount of docetaxel (w/w), the particle size can become larger, which typically results in the formulation becoming cloudy. Accordingly, in some embodiments, the amount of oil is less than about three times the amount of docetaxel (w/w), or equal to or less than about 2.5 times the amount of docetaxel (w/w), or equal to or less than about 2 times the amount of docetaxel (w/w), or equal to or less than about 1.5 time the amount of docetaxel (w/w), or is about equal to the amount of docetaxel (w/w), or is about half of the amount of docetaxel (w/w).

Also present in the nanodispersion formulations is a non-ionic surfactant, which may include one or more non-ionic surfactants. Surfactants of interest include any type of non-ionic surfactant that can be used for pharmaceutical formulations. Non-ionic surfactants of interest include, but are not limited to, polyoxyalkylene copolymer, and sorbitan fatty acid esters. In some embodiments, the sorbitan fatty acid ester is a polyoxyethylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan tristearate (Tween® 65); polyoxyethylene sorbitan trioleate (Tween® 85); polyethylene glycol 400 monostearate; polysorbate 60; (Tween® 60); polyoxyethylene monostearate (Myrj® 49); polysorbate 80 (Tween® 80); polysorbate 40 (Tween® 40); and polysorbate 20 (Tween 20)) or sorbitan fatty acid esters (e.g., sorbitan trioleate (Span® 85); sorbitan tristearate (Span® 65); sorbitan sesquioleate (Arlacel® 83); glyceryl monostearate; sorbitan monooleate (Span® 80); sorbitan monostearate (Span® 60); sorbitan monopalmitate (Span® 40); and sorbitan monolaurate (Span® 20)). In some embodiments, the non-ionic surfactant is polysorbate 80. In some embodiments, the polysorbate 80 is refined grade.

The amount of non-ionic surfactant in the nanodispersion formulation may vary. In some instances, the amount of non-ionic surfactant in the nanodispersion formulation is about 40 wt % or more. In some instances, the non-ionic surfactant is present in an amount ranging from about 40 to about 75 wt %, or from about 50 to about 65 wt %, or from about 50 to about 60 wt %, or from about 50 to about 57 wt %, or from about 57 to about 65 wt %. The combination ratio by weight of the oil and the surfactant in the subject nanodispersion formulations may vary, and is in some instances about 1/100, or about 1/50, or about 1/40, or about 1/30, or about 1/20, or about 1/10, or about 1/8, or about 1/6, or about 1/4, or about 1/2, or about 1/1.

In some instances, nanodispersion formulations of the invention further include a non-aqueous solvent, which may include one or more non-aqueous solvents. Non-aqueous solvents of interest include, but are not limited to, propylene glycol, polypropylene glycol, polyethylene glycol (such as PEG 300, PEG 400, PEG 600, PEG 800, PEG 1000, etc., where in certain embodiments polyethylene glycols, when employed, have an average molecular weight of 1000 or less), glycerin, triacetin, dimethyl isosorbide, glycofurol, propylene carbonate, dimethyl acetamide or a mixture thereof. In some embodiments, the non-aqueous solvent is polyethylene glycol. In some embodiments, the non-aqueous solvent is polyethylene glycol 400. In some embodiments, the polyethylene glycol has a melting point of less than 0° C. In some embodiments, the non-aqueous solvent is polyethylene glycol 300.

The non-aqueous solvent may be present in varying amounts, and in some instances is present in an amount ranging from about 20 to about 60 wt %, or from about 35 to about 45 wt %, including amounts ranging from about 35 to about 40 wt %, or from about 40 to about 45 wt %. The combination ratio by weight of the oil and the non-aqueous solvent in the nanodispersion formulations may vary, and in some instances is about 1/100, or about 1/50, or about 1/40, or about 1/30, or about 1/20, or about 1/10, or about 1/7, or about 1/5, or about 1/3, or about 1/1.

In some instances, the amount of non-aqueous solvent is determined based on the amount of non-ionic surfactant in the formulation. In such cases, the amount of non-ionic surfactant (w/w) should be about equal to or greater than the amount of non-aqueous solvent (w/w) in the formulation, or greater than about 95% of the amount of non-aqueous solvent (w/w), or in other words, the ratio by weight of the non-ionic surfactant and the non-aqueous solvent is about 0.95 or more. As is shown in the Examples, when used in such amounts, the stability of the formulation is enhanced.

The nanodispersion formulations disclosed herein further comprise an organic acid component. The organic acid as used herein is soluble in the non-aqueous, ethanol-free formulation (in the amount used) and does not contain any significant amount of (i.e., is substantially-free of) its conjugate base (or salt thereof). In some embodiments, the organic acid contains less than about 5% of its conjugate base (or salt thereof), or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1%, or less than about 0.5%, or less than about 0.1%, or less than about 0.05%, or less than about 0.01%. Organic acids of interest upon which the organic acid component may be based include lactic acid, succinic acid, malic acid, tartaric acid and acetic acid. In some embodiments, the nanodispersion formulations disclosed herein do not contain a buffer or a mineral acid. In some embodiments, the organic acid component is lactic acid. The lactic acid can be D- or L-lactic acid, or a mixture thereof.

The organic acid may be present in varying amounts, and in some instances is present in an amount ranging from about 0.3 to about 3 wt %, or from about 0.3 to about 1 wt %, or from about 0.3 to about 0.6 wt %, or from about 0.4 to about 0.5 wt %, or from about 0.5 to about 1 wt %, or from about 0.7 to about 1 wt %, or about 0.8 wt %.

In some embodiments, the present disclosure provides for a non-aqueous, ethanol-free docetaxel liquid nanodispersion formulation comprising: docetaxel, soybean oil in an amount ranging from about 1 to about 5 wt %, polysorbate 80 in an amount ranging from about 50 to about 60 wt %, polyethylene glycol in an amount ranging from about 35 to about 45 wt %, and lactic acid in an amount ranging from about 0.3 to about 1% w/w and wherein said lactic acid is substantially lactate-free.

In another embodiment, the present disclosure provides for a non-aqueous, ethanol-free docetaxel liquid nanodispersion formulation comprising: docetaxel, a medium chain triglyceride in an amount ranging from about 1 to about 5 wt %, polysorbate 80 in an amount ranging from about 50 to about 60 wt %, polyethylene glycol in an amount ranging from about 35 to about 45 wt %, and lactic acid in an amount ranging from about 0.3 to about 1% w/w and wherein said lactic acid is substantially lactate-free.

The nanodispersion formulation typically has a pH of less than about 3.5, or less than about 3.4, or less than about 3.3, or less than about 3.25, or less than about 3.2, or less than about 3.1, or from about 3.0 to about 3.1.

Methods of Preparing Docetaxel Nanodispersion Formulations

Nanodispersion formulations may be prepared according to any convenient protocol. As such, the components of the desired nanodispersion may be combined under conditions sufficient to produce the desired nanodispersion. Accordingly, an amount of docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base, may be combined under conditions sufficient to produce a nanodispersion. The components may be combined in any convenient order. The components may be combined at any convenient temperature, e.g., room temperature or elevated temperatures, such as temperatures ranging from about 30 to about 95° C., about 50 to about 60° C., or about 70 to about 80° C. Certain of the components may be combined with each other, and then combined together, or all of the components may be combined at substantially the same time. Combination may include various manners of agitation or mixing, e.g., stirring, sonication, etc., in order to produce the desired nanodispersion.

As indicated above, an initial nanodispersion is prepared that includes docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base. In certain embodiments, the initial nanodispersion is clear. By clear is meant that the nanodispersion is a translucent, if not transparent liquid, i.e., the liquid is pellucid. As such, the initial preparation is not cloudy. Further details regarding the nanodispersions that may be prepared from the docetaxel initial composition precursors are provided below.

In some embodiments, the present disclosure provides a method for making the non-aqueous, ethanol-free docetaxel nanodispersion formulation as disclosed herein, the method comprising combining docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid; and sterilizing the product thereof to provide the non-aqueous, ethanol-free docetaxel nanodispersion formulation.

The preparation methods can be carried out at certain temperature such as temperatures ranging from about 30° C. to about 95° C., about 50° C. to about 60° C., about 60° C. to about 70° C., or about 70° C. to about 80° C. Specific examples are provided in the Experimental section, below.

In some embodiments, the method for making the non-aqueous, ethanol-free docetaxel nanodispersion formulation as disclosed herein does not comprise the addition of water and/or ethanol, followed by removal thereof. Accordingly, since the non-aqueous, ethanol-free docetaxel nanodispersion formulation has not been contacted with either water or ethanol during the formulation process, it is substantially free of water (i.e, less than about 3%) and free of ethanol. In some embodiments, the formulation is essentially free of water (i.e, less than about 1%). In addition, in certain embodiments, the formulations do not have any detectable amount of ethanol. This results in a formulation which is stable and can be suitable for subjects having an adverse reaction to ethanol. In some embodiments, the formulations do not contain activated carbon.

Where desired, an amount of the nanodispersion may be loaded into an individual dosage container, e.g., vial, which holds the nanodispersion and keeps it sterile during shipping, storage, and handling. Before or during the loading stage, the nanodispersion can be passed through a sub-micron sterilizing filter, e.g., a 0.2 μm hydrophilic filter) which has a sufficiently small pore size to remove any bacteria or viruses. The sterilizing filters of interest include, but are not limited to hydrophilic filters. In some embodiments, the filter could be a CA (Cellulose Acetate) membrane filter, PTFE (Polytetrafluoroethylene) membrane filter, PVDF (Polyvinylidene fluoride or polyvinylidene difluoride) membrane filter or PES (Polyethersulfone) membrane filter.

As used herein, the term “vial” refers to any container that is used to hold the nanodispersion formulation. Many pharmaceutical vials are made of clear glass, which allows several advantages, including visual inspection of the enclosed drug (to ensure that it is still in a clean, non-caramelized, non-collapsed form, when it is ready for use) and of the container itself (to ensure that it does not have a hairline crack in one of the walls, which could jeopardize or destroy sterility of the enclosed drug). Various types of pharmaceutical vials are known. Single-chamber vials can be sealed with rubber or plastic plugs that will allow a hypodermic needle to be pushed through the rubber seal. Examples include any hydrolytically stable glass, such as a borosilicate Type I or soda-lime-silica Type II glass vial, having a suitable stopper (e.g., Teflon coating). Alternately, a single-chamber vial can be made of a brittle and easily breakable material, inside a sealed bag that can contain an aqueous solution (such as physiological saline or a dextrose solution, in an intravenous infusion bag); if this type of vial is broken, it will release its contents into the still-sealed bag, for mixing. In yet other embodiments, two-chamber vials or analogous structures, e.g., as described in Published United States Application Publication No. 2003/0099674 and U.S. Pat. No. 4,781,354 may be employed. Other methods for preparing the docetaxel formulations disclosed herein can be found in Published United States Application Publication No. 2011/0269829, the entirety of which is incorporated herein by reference.

Docetaxel Formulations and Methods of Use

Following preparation of the nanodispersion formulation, e.g., as described above, at the time of desired administration to a subject, a dosage amount of the nanodispersion may be combined with an aqueous medium to prepare a docetaxel diluted solution that is suitable for use. The dosage amount of the nanodispersion formulation may be combined with any suitable aqueous medium, where aqueous mediums of interest include, but are not limited to: deionized water, USP water for injection (WFI), salines, transfusion solutions, physiological solutions, etc. In some embodiments, the aqueous medium comprises an aqueous 0.9% sodium chloride solution or an aqueous 5% dextrose solution. The liquids to nanodispersion (high viscous liquid) ratio employed during preparation of the diluted solution may vary, and in certain embodiments ranges from 0.5 to 300, such as 1 to 100, 2 to 50 or 2 to 20, and including 2 to 10. In some instances, the dosage amount of nanodispersion formulation that is combined with the aqueous medium ranges from 100 to 1200 g, such as 300 to 600 g and the amount of aqueous medium that is combined with the dosage amount ranges from 100 to 1200 ml, such as 250 to 600 ml.

The docetaxel diluted solutions prepared from the nanodispersion formulations are liquid preparations that are a suspension of small particles (i.e., globules) of one liquid in a second liquid with which the first liquid will not mix. The water present in the docetaxel diluted solutions may be any convenient water, including deionized water, USP water for injection (WFI), etc.

The docetaxel diluted solutions include docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and an organic acid, wherein said organic acid is soluble in the non-aqueous solvent and is substantially free of any conjugate base, and an aqueous medium. In certain embodiments, the docetaxel diluted solutions are clear. By clear is meant that the diluted solution is a translucent, if not transparent liquid, i.e., the liquid is pellucid. As such, the diluted solution is not cloudy, e.g., as a suspension may appear. Further details regarding the docetaxel diluted solutions that may be prepared from the docetaxel nanodispersion precursors are provided below. In some instances, the particle size of the final docetaxel diluted solutions ranges from about 3 to about 70 nm, such as about 5 to about 50 nm and including about 7 to about 30 nm, such as about 8 to about 15 nm. In some instances, any difference in particle size between the non-aqueous formulation and the docetaxel diluted solution is minimal, such that the particle sizes in the non-aqueous formulation and the docetaxel diluted solution are substantially the same. In some instances, any difference in particle size between the non-aqueous formulation and the docetaxel diluted solution is about 30 nm or less, such as about 20 nm or less, about 15 nm or less, about 10 nm or less, or about 5 nm or less. Without wishing to be bound by any one theory, it is contemplated that the inclusion of the oil in the nanodispersion formulations helps stabilize the diluted solution.

Where desired, the nanodispersion formulation may be stored for a period of time prior to combination with the aqueous medium. This storage time of the nanodispersion composition may vary, where storage times may be about 1 year or more, such as about 2 years or more, including about 3 years or more. While the storage conditions may vary, in certain instances the storage conditions are characterized by a temperature ranging from about 5 to about 60° C., e.g., about 5° C., such as about 8 to about 40° C., e.g., about 25° C. The activity of the docetaxel is substantially preserved during the storage period, such that the nanodispersion formulations are storage stable. As such, the activity of the docetaxel in the docetaxel diluted solution following storage is substantially the same as that in the nanodispersion prior to being combined with an aqueous medium, where the magnitude of any difference in activity between the non-aqueous formulation and the docetaxel diluted solution may be about 15% or less, such as about 10% or less, including about 5% or less.

As shown in Example 2, the nanodispersion formulation according the present disclosure is stable for more than about 3 months at about 40° C., or is stable for more than about 6 months at about 40° C. (see Example 3). In contrast, the formulations which contain lactic acid buffer (i.e., lactic acid with sodium lactate) rather than lactic acid alone show signs of deterioration during the same time period. The stability of the present nanodispersion formulations can be determined by methods known in the art, such as by measuring the percent deterioration of the docetaxel peak by HPLC or by measuring recovery rate for the docetaxel by HPLC. In some embodiments, the nanodispersion formulation exhibits a docetaxel recovery rate of more than about 95% after 6 months at about 40° C., or about 96% or more, or more than about 97% or more, or more than about 98% or more, or about 99% or more. The recovery rate is calculated based on the measured amount of docetaxel in the formulation, which is not necessarily the amount of docetaxel added to the formulation (thus eliminating any potential error from an impurity in the docetaxel).

The docetaxel diluted solutions that are produced upon dilution of the nanodispersion formulation with the aqueous medium can have a physiologically acceptable pH. In certain embodiments, the pH of the diluted solutions ranges from about 2.5 to about 8, such as from about 3 to about 7, including from about 3.5 to about 6. The docetaxel diluted solutions are clear (i.e., transparent) formulations. The concentration of docetaxel in the diluted solution may vary, ranging in some embodiments from about 0.05 to about 10 mg/ml, such as about 0.2 to about 3 mg/ml.

The combination protocol may vary, where agitation may be employed, e.g., by stirring, by kneading a bag that includes both the nanodispersion formulation and the aqueous medium.

Methods of using the docetaxel diluted solutions include administering an effective amount of the docetaxel diluted solution to a subject in order to treat the subject for a target condition of interest. By “treating” or “treatment” is meant at least a suppression or an amelioration of the symptoms associated with the condition afflicting the subject, where suppression and amelioration are used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g., symptom, associated with the condition being treated, such as pain. As such, treatment also includes situations where the condition is completely inhibited, e.g., prevented from happening, or stopped, e.g., terminated, such that the subject no longer experiences the condition. As such, treatment includes both preventing and managing a condition.

In practicing the methods, the diluted solutions disclosed herein can be parenterally administered to a subject. By “parenteral administration” is meant administration by a protocol that delivers a quantity of the diluted solutions to the subject, e.g., a patient suffering from a cellular proliferative disease, by a route other than the digestive tract. Examples of parenteral administration include, but are not limited to, intramuscular injection, intravenous injection, and the like. In certain embodiments, parenteral administration is by injection using an injection delivery device. The amount of diluted solutions that is administered to the subject may vary depending on a number of factors, such as patient specifics, nature of condition, etc.

In certain embodiments, the subject methods include a diagnostic step. Individuals may be diagnosed as being in need of the subject methods using any convenient protocol. In addition, individuals may be known to be in need of the subject methods, e.g., they are suffering from a target disease condition (e.g., cellular proliferative disease, prior to practicing the subject methods). Diagnosis or assessment of target condition can be performed using any convenient diagnostic protocol.

Methods of the invention may further include assessing the efficacy of the treatment protocol that includes administration of the docetaxel diluted solution. Assessing the efficacy of treatment may be performed using any convenient protocol.

The docetaxel diluted solutions of the invention may be administered to a variety of different types of subjects. Subjects of interest include, but are not limited to: mammals, both human and non-human, including the orders carnivore (e.g., dogs and cats), rodentia (e.g., mice, guinea pigs, and rats), lagomorpha (e.g., rabbits) and primates (e.g., humans, chimpanzees, and monkeys). In certain embodiments, the subjects, e.g., patients, are humans.

Accordingly, provided herein are methods of administering an ethanol-free docetaxel liquid nanodispersion formulation to a subject, the method comprising: (a) combining the ethanol-free docetaxel nanodispersion formulation according to the present disclosure with an aqueous medium to provide an ethanol-free docetaxel diluted solution; and (b) administering the ethanol-free docetaxel diluted solution to the subject.

In some embodiments, the method of using the docetaxel nanodispersion formulation comprises the steps of: (a) aseptically withdrawing the desired amount of the docetaxel nanodispersion formulation (such as a formulation comprising about 20 mg docetaxel/mL) with a calibrated syringe, (b) injecting said formulation into a 250 mL infusion bag or bottle of either 0.9% sodium chloride solution or 5% dextrose solution to provide a diluted solution having a final docetaxel concentration of from about 0.3 mg/mL to about 0.74 mg/mL, and (c) administering said diluted solution to a patient. If a dose greater than 200 mg of docetaxel is required, one may use a larger volume of the infusion vehicle so that a concentration of 0.74 mg/mL docetaxel is not exceeded.

In certain embodiments, the volume of diluted solution that is administered to a subject may range from about 100 to about 1000 ml, such as about 200 to about 600 ml. The time period over which this volume is administered may vary, ranging from about 0.5 to about 6 hr, such as from about 1 to about 3 hr. Dosages administered to a subject during a given procedure may also vary, ranging in some instances from about 20 to about 500 mg/m², such as from about 50 to about 300 mg/m².

In determining whether to administer the diluted solution to a particular given subject, care will be taken to ensure that the formulation is not contraindicated for that subject. As such, symptoms of the subject may be assessed to ensure that administration of the diluted solution will not have adverse effects that outweigh any benefit that the diluted solution may provide.

Utility

The subject diluted solutions and methods find use in a variety of applications, including the treatment of subjects suffering from cellular proliferative disease conditions. Cellular proliferative diseases that may be treated with formulations of the invention include, but are not limited to: carcinomas, myelomas, neuroblastomas, or sarcomas, of the brain, breast, lung, colon, prostate or ovaries, as well as leukemias or lymphomas. Specific disease conditions of interest include, but are not limited to, human ovarian cancer, breast cancer, malignant lymphoma, lung cancer, melanoma, and Kaposi's sarcoma.

Kits

Also provided are kits that find use in practicing the subject methods, as described above. For example, kits for practicing the subject methods may include a quantity of the nanodispersion formulation, present in unit dosages, e.g., vials, or a multi-dosage format. As such, in certain embodiments, the kits may include one or more unit dosages (e.g., vials) of the nanodispersion formulation. The term “unit dosage”, as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of the subject nanodispersion formulation calculated in an amount sufficient to produce the desired effect. The amount of the unit dosage of the subject formulation depends on various factors, such as the particular active agent employed, the effect to be achieved, and the pharmacodynamics associated with the active agent in the subject. In yet other embodiments, the kits may include a single multi-dosage amount of the formulation.

In certain embodiments, the kits may further include an amount of an aqueous medium suitable for use in production of the docetaxel diluted solution. The aqueous medium may be any convenient aqueous medium, such as described above, present in any suitable container, e.g., an IV bag.

In some embodiments, the kits may include a syringe which is suitable to prepare the docetaxel diluted solution. A syringe with graduations is preferred to measure a certain amount of the docetaxel nanodispersion.

In addition to the above components, the subject kits may further include instructions for practicing the subject methods. These instructions may be present in the subject kits in a variety of forms, one or more of which may be present in the kit. One form in which these instructions may be present is as printed information on a suitable medium or substrate, e.g., one or more pieces of paper on which the information is printed, in the packaging of the kit, in a package insert, etc. The instructions may be present on a computer readable medium, e.g., diskette, CD, DVD, etc., on which the information has been recorded. The instructions may be present on a website, which may be used via the internet to access the information at a removed site. Other convenient means are possible and may be included in the kits.

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

EXPERIMENTAL Example 1: General Formulation

Nanodispersion Formulation

Taxane (e.g., anhydrous docetaxel or docetaxel trihydrate), oil (e.g., soybean oil), surfactant (e.g., polysorbate 80), non-aqueous solvent (e.g., polyethylene glycol 300, average molecule weight=300) and organic acid component (e.g., lactic acid or lactic acid mixture of 4:1 by wt lactic acid to sodium lactate) are placed into a beaker. The beaker is heated to 70-80° C. and the ingredients mixed via ultrasonic dispersion.

The resultant solution is poured into a vial through a 0.2 micron filter while applying nitrogen and the tube sealed. Optionally, steam treatment (95° C.×30 min.) can then be applied.

Diluted Solution

The resultant nanodispersion formulation is then placed into a test tube. Purified water was added and the tube was shaken by hand for about 20 seconds to obtain a clear solution. The particle size is then measured via a particle size distribution in the dynamic light scattering measurement protocol.

The various studies performed using nanodispersion formulations and diluted solutions prepared as described above are outlined in the Examples below.

Example 2: Oil Studies

Nanodispersion samples were prepared according to Example 1 and 0.5 ml of the sample was added to 25 ml of 5% glucose solution in a glass centrifuge tube. The tube was shaken by hand for about 20 seconds. The diluted sample was evaluated by visual inspection for phase separation, color, clarity, consistency and particulates in the clear, glass tube.

Oil Study 1

In the following Example, all components were mixed, water was added, and the formulation was autoclaved then left at room temperature for four days. As shown in Table 1, when the ratio of soybean oil to docetaxel (by weight) is 1 (Entry (1)) or 2.5 (Entry (2)), there was no phase separation observed after 4 days.

TABLE 1 Formulation A (1) (2) Docetaxel 4 4 Soybean oil 4 10 Polysorbate 80 (GS) 100 100 Polyglycol (PG) 90 90 Glycerin 10 10 LA-Buffer (8:2) 1 1 Physical appearance ∘ ∘ of the dispersion sample (4 days later) ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation Oil Study 2

As shown in Table 2, when the ratio of soybean oil to docetaxel (by weight) is three (Entry (2)), the quality of the nanodispersion declines.

TABLE 2 Formulation B (1) (2) Docetaxel trihydrate 4 4 MCT (NEOBEE 1053) 8 12 Polysorbate 80 95 95 (TO-10MV) Lactic Acid Buffer 1 1 (LA:70% LANa = 8:2) PEG 300 (MG300) 70 70 Physical appearance ∘ x* of dispersion ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation *gelatinous look Oil Study 3

The following docetaxel nanodispersion solutions were evaluated by visual inspection for phase separation, color, clarity, consistency and particulates in clear, glass tubes when they were prepared. In addition, 5.25 ml of each sample was combined with 250 ml of 0.9% sodium chloride solution or 5% glucose solution. The diluted solution was then visually inspected after 6 hours and 24 hours. The results are shown in Tables 3 and 4. The stability of the docetaxel diluted solution was increased by adding oil.

TABLE 3 Formulation C (1) (2) (3) (4) Docetaxel 4 4 4 4 MCT (NEOBEE 1053) 0 0 4 4 Polysorbate 80 100 100 100 100 (TO-10MV) PEG 300 (MG300) 40 40 40 40 PEG 400 (MG400) 10 60 10 60 Lactic Acid Buffer 1 1 1 1 Physical appearance ∘ ∘ ∘ ∘ of dispersion (0 days) Physical appearance ∘~Δ ∘~Δ ∘ ∘ of diluted solution (6 hrs) Physical appearance x x Δ Δ of diluted solution (24 hrs) ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

TABLE 4 Formulation D (1) (4) Docetaxel trihyrate 4 4 Soybean oil 0 4 Polysorbate 80 95 95 (TO-10MV) Lactic Acid Buffer 1 1 LA:70% LANa = 8:2) PEG 300 (MG300) 70 70 Physical appearance x x ∘ Δ of diluted solution (6 hrs) ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation Oil Study 4: Comparison Example

Table 5 shows the stability of formulations provided using water in the formulation process (see US2011/0269829, section II: Docetaxel Formulations, A. Working example 1). Particle size was measured before the water removal process.

TABLE 5 Formulation E Formulation E Formulation F Formulation F (1) (3) (1) (3) Docetaxel 4 (200 mg) 4 (200 mg) 4 (200 mg) 4 (200 mg) Soybean oil 10 (500 mg) 4 (200 mg) — — MCT* — — 10 (500 mg) 4 (200 mg) Polysorbate 80 100 (5 g) 100 (5 g) 100 (5 g) 100 (5 g) PG 100 (5 g) 100 (5 g) 100 (5 g) 100 (5 g) Lactic Acid Buffer 5 5 5 5 (LA 155 mg, 70% LANa 95 mg) Particle size (nm) 17.5 (wide) 12.8 14.7 (wide) 12.1 Physical appearance Cloudy, phase Transparent, Transparent, Transparent, of dispersion after separation clear clear clear water removal (0 days) *MCT: Nisshin O.D.O

Example 3: PEG (Polyethylene Glycol)

Nanodispersion samples were prepared according to Example 1 and evaluated by visual inspection for phase separation, color, clarity, consistency and particulates in clear, glass vials.

PEG Study 1: Stability

The formulations shown in Table 6 were stored at 40° C./75% RH for 6 months (Formulation G, Entries (1), (2), and (3)) and 5° C. for 6 months (Formulation H, Entries (1), (2), and (3)). Table 6 shows that the formulations which used PEG 300, which has a melting point of −15° C. to −8° C., maintained transparency.

TABLE 6 Formulation G Formulation H (1)(2)(3) (1)(2)(3) Docetaxel trihydrate 4 mg 4 mg MCT (NEOBEE 1053)  4  4 Polysorbate 80 95 95 (TO-10MV) PEG 300 (MG300) 70 — PEG 400 (MG400) — 70 LA-Buffer  1  1 (LA:70% LANa = 8:2) Physical appearance (1)(2)(3) (1)Yellow/ of dispersion after Yellow/ Transparent/ 6 months at 40° C. Transparent/ Clear Clear (2)(3) Yellow/ Transparent/ Slightly cloudy Physical appearance (1)(2)(3) (1)(2)(3) of dispersion after Yellow and cloudy. Yellow and cloudy. 6 months at 5° C. Floating subjects Floating subjects observed when were seen when temperature the temperature was still low. was still low. When the When the temperature was temperature was back to room back to room temperature, it temperature, it was Yellow/ was Yellow/ Transparent/ Transparent but Clear. still Cloudy and Ununiform. PEG Study 2: Solubility

Formulations of anhydrous docetaxel and either PEG 400 or PEG 300 were prepared and mixed using ultrasonic dispersion for 5 minutes at 70-80° C. 1 g of PEG 400 dissolved 25 mg of anhydrous docetaxel. 1 g of PEG 400 did not fully dissolve 50 mg of anhydrous docetaxel. 1 g of PEG 300 dissolved 100 mg of anhydrous docetaxel. Accordingly, PEG 300 was able to better solubilize anhydrous docetaxel.

PEG Study 3: Polysorbate/PEG Ratio

As shown in Table 7, when the ratio of polysorbate to PEG is greater than or equal to 1 (i.e., the amount of polysorbate is greater than or equal to the amount of PEG), the formulations exhibit stability compared to formulations having less PEG with respect to polysorbate.

TABLE 7 Formulation Formulation Formulation Formulation Formulation Formulation C (4) C (5) C (6) I (4) I (5) I (6) Docetaxel 4 4 4 Docetaxel trihydrate 4 4 4 MCT (NEOBEE 4 4 4 4 4 4 1053) Polysorbate 80 (TO- 100 100 100 95 95 95 10MV) PEG300 (MG300) 40 40 20 100 50 0 PEG400 (MG400) 10 60 80 0 50 100 LA-Buffer 1 1 1 1 1 1 Total 159 209 209 204 204 204 Ratio 2 1 1 0.95 0.95 0.95 Polysorbate/PEG Physical appearance ∘ ∘ ∘ x Slightly x Slightly ∘ of dispersion after 0 cloudy Cloudy day Physical appearance ∘ x phase x phase x phase x phase x phase of dispersion after 1 separation separation separation separation separation day slightly Physical appearance x x x phase of dispersion after 6 separation days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation PEG Study 4: Polysorbate/PEG Ratio (No API)

In Tables 8, 9 and 10, when the ratio of polysorbate to PEG is greater than or equal to 0.95 or 1 (i.e., the amount of polysorbate is greater than or about equal to the amount of PEG), the formulations exhibit stability compared to formulations having more PEG with respect to polysorbate (no taxane was added in Tables 8 and 9).

TABLE 8 Formulation J (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) Ratio 2 1 0.95 0.8 0.33 1.33 4 Polysorbate/PEG Polysorbate 80 (TO- 100 100  100  95 95 95 8 8 80 80 10MV) PEG 300 (MG300) 40 40 20 100  50  0 10 24 0 0 PEG 400 (MG400) 10 60 80  0 50 100  0 0 60 20 Physical appearance ∘ ∘ ∘ ∘ ∘ Δ x x ∘ ∘ of dispersion after 0 day Physical appearance ∘ ∘ ∘ x x x x x ∘ ∘ of dispersion after 5 days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

TABLE 9 Formulation K 1 2 3 4 5 6 7 8 9 10 11 Ratio 0.33 0.67 1 Polysorbate/PEG PEG 300/400 0/100 25/75 50/50 75/25 100/0 0/100 25/75 50/50 75/25 100/0 75/25 (MG300/400) Polysorbate 80 8 8  8 8 8 16 16 16 16 16 16  (TO-10MV) PEG 300 0 6 12 18  24   0  6 12 18 24 12  (MG300) PEG 400 24  18  12 6 0 24 18 12  6  0 4 (MG400) Physical x x x x x x x x x x ∘ appearance of dispersion after 0 day ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

TABLE 10 Formulation L (1) (2) (3) (4) Docetaxel trihydrate 4 4 4 4 MCT (NEOBEE 1053) 0 0 4 4 Polysorbate 80 95 95 95 95 (TO-10MV) PEG 300 (MG300) 40 85 40 85 PEG 400 (MG400) 10 0 10 0 Lactic Acid Buffer 1 1 1 1 Total 150 185 154 189 Polysorbate/PEG 1.9 1.12 1.9 1.12 Ratio Physical appearance ∘ ∘ ∘ ∘ of dispersion after 0 day Physical appearance ∘ ∘ ∘ ∘ of dispersion after 7 days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation PEG Study 5: Polysorbate/PEG Ratio

Tables 11 and 12 show that the oil contributes to the stability of the docetaxel nanodispersion.

TABLE 11 Formulation M (1) Docetaxel trihydrate 4 MCT (NEOBEE 1053) 4 Polysorbate 80 95 (TO-10MV) PEG 300 (MG300) 70 Lactic Acid Buffer 1 Total 174 Polysorbate/PEG 1.36 Ratio Physical appearance ∘ of dispersion after 5 days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/ precipitation

TABLE 12 Formulation N (4) (6) Docetaxel trihydrate 4 4 MCT (NEOBEE 1053) 0 0 Polysorbate 80 95 95 (TO-10MV) PEG 300 (MG300) 70 85 Lactic Acid Buffer 1 1 Polysorbate/PEG 1.36 1.12 Ratio Physical appearance ∘ Δ of dispersion after 0 days Physical appearance of ∘ ∘ dispersion after 1 days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation PEG Study 6: PEG and PG (Polyglycol) Comparison

Table 13 shows that even with a polysorbate/PG ratio greater than 1, the formulation did not satisfy the desired quality level.

TABLE 13 Formulation O (1) Docetaxel trihydrate 4 (120 mg) MCT (NEOBEE 1053) 4 (120 mg) Polysorbate 80 95 (2.85 g) (TO-10MV) Lactic Acid Buffer 1 (30 mg) (LA:70% LANa = 8:2) Polyglycol 70 70 (2.1 g) Polysorbate/Poly- 1.36 glycol Ratio Physical appearance Δ of dispersion after 0 days Physical appearance x of dispersion after 1 days ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

Table 14 shows that even when the polysorbate/polyglycol&PEG ratio was over 1, when polyglycol was added, the quality of the nanodispersion declined.

TABLE 14 Formulation P (1) (2) Docetaxel trihydrate 4 (120 mg) 4 (120 mg) MCT (NEOBEE 1053) 4 (120 mg) 4 (120 mg) Polysorbate 80 95 (2.85 g) 95 (2.85 g) (TO-10MV) Lactic Acid Buffer 1 (30 mg) 1 (30 mg) (LA:70% LANa = 8:2) PEG 300 (MG300) 40 (1.2 g) 30 (900 mg) Polyglycol 0   40 (1.2 g) Polysorbate/ 0.42 1.36 Polyglycol&PEG Ratio Physical appearance Slightly Cloudy, of dispersion after cloudy Particulates 2 days Physical appearance ∘, Δ N/A of diluted solution (2 samples) (3 hr) ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

Example 5: Buffering Agent

Lactic Acid Study 1: Lactic Acid and Lactic Acid Buffer

Lactic acid lowered the pH further than lactic acid buffer (docetaxel is stable at pH 3.0-4.0) (Table 15).

TABLE 15 Formulation Q (1) (2) (3) (4) Docetaxel — 4 (120 mg) — — Docetaxel trihydrate 4 (120 mg) — — — MCT (NEOBEE 1053) 4 (120 mg) 4 (120 mg) 4 (200 mg) 4 (200 mg) Polysorbate 80 95 (2.85 g) 95 (2.85 g) 95 (4.75 g) 95 (4.75 g) (TO-10MV) PEG 300 (MG300) 70 (2.1 g) 70 (2.1 g) 70 (3.5 g) 70 (3.5 g) Lactic Acid Buffer — — — 1 (50 mg) (LA:90% LANa = 8:2) Lactic Acid 0.8 (24 mg) 0.8 (24 mg) 0.8 (40 mg) — Physical appearance ∘ two ∘ two ∘ ∘ of dispersion bubbles were bubbles were seen seen pH of diluted 3.19 3.16 3.15 3.43 solution ∘: Clear and free from particle matter Δ: Very slightly cloudy x: Slightly cloudy/cloudy/phase separation/precipitation

Example 6: Water Free Formulation

Lactic Acid Study 1: LA and LA Buffer

The formulations in Tables 16 and 17 exhibit stability over a six month period. In the Tables, the lactic acid formulations exhibited an enhanced stability when compared to the lactic acid buffer formulations (Table 16 vs Table 17).

TABLE 16 Formulation Formulation Formulation Formulation Formulation Formulation R S (1)(2) G (1)-(3) T U (2) V Docetaxel — — — — — 4 Docetaxel 4 4 4 4 4 — trihydrate NEOBEE 1053 4 4 4 4 4 4 Polysorbate 80 95  95  95  95  — — (TO-10MV) Polysorbate 80 — — — — 95 95 (GS) PEG 300 (MG300) 70  70  35 + 35 35 + 35 70 70 Lactic Acid Buffer 1 1 (100 mg) 1 (100 mg) — — — (LA:70% LANa = 8:2) Lactic Acid Buffer — — — 1 — — (LA:90% LANa = 8:2) Dehydrated Lactic — — — — 0.8 0.8 Acid Buffer (LA:90% LANa = 8.4:1.6) Docetaxel recovery 86.7-89.3% 86.7-89.3% 86.4-88.2% (1 month) 94.5% 91.1% rate after 6 months 95.8% Degradation peak  3.3-3.6%  3.3-3.6%  2.6-2.9% (1 month) 1.9% 3.9% after 6 months 1.7% pH of docetaxel   3.50   3.50 3.61-3.62 (1 month) 3.55 3.54 nanodispersion 3.53 pH of diluted 3.69-3.70 3.69-3.70 3.77-3.80 — — — solution

TABLE 17 Formulation W Formulation X Formulation Y Formulation Z (1)-(3) (1)-(3) (1)-(3) (1)-(3) Docetaxel  4 —  4 — Docetaxel trihydrate —  4 —    4.27 NEOBEE 1053  4  4 — — Soybean oil — —  4  4 Polysorbate 80 (GS) 95 95 95 95 PEG 300 (MG300) 70 70 70 70 Lactic Acid   0.8   0.8   0.8   0.8 Docetaxel recovery 96.9-98.0%    99.3-100.7%  101.6-102.3%  101.4-102.4% rate after 6M Degradation peak 1.0-1.4% 0.8% 0.1% 0.1% after 6 months pH of docetaxel 3.07-3.10  3.01-3.02 3.02-3.06 3.05-3.11 nanodispersion pH of diluted — 3.54-3.57 3.54-3.57 3.54-3.57 solution

Example 1a: Docetaxel Nanodispersion Formulations Using Lactic Acid

Docetaxel, an oil, a non-ionic surfactant, a non-aqueous solvent and lactic acid were placed into a beaker. The beaker was heated to 70-80° C., the ingredients mixed via ultrasonic dispersion, the resultant solution was added to a vial tube through a 0.2 μm filter while applying nitrogen and the tube was sealed.

To determine particle size, a non-aqueous composition as prepared using the general procedure outlined above was placed into a test tube. Purified water was added and the tube was shaken by hand for about 20 seconds to obtain a clear solution. When the particle size was measured via a particle size distribution in the dynamic light scattering measurement protocol, the average particle size was observed to be between about 8 and about 13 nm.

Specific formulations (in mg) as prepared via the general method disclosed above are summarized in Table 1a, below.

TABLE 1a Oil MCT Surfactant Solvent Docetaxel (NEOBEE ® MCT Soybean Polysorbate PEG 300 Lactic No. Anhydrous Trihydrate #1053) (Panacet ®) Oil 80 (MG-300) Acid 1A 4 4 95 70 0.8 1B 4 4 95 70 0.8 1C 4 4 95 70 0.8 2A 4 4 95 70 0.8 2B 4 4 95 70 0.8 2C 4 4 95 70 0.8 3A 4 4 95 70 0.8 3B 4 4 95 70 0.8 4A 4.27* 4 95 70 0.8 4B 4.27* 4 95 70 0.8 4C 4.27* 4 95 70 0.8 5A 4 4 95 70 0.8 5B 4 4 95 70 0.8 5C 4 4 95 70 0.8 6 4 4 95 70 0.8 *4.27 mg Docetaxel trihydrate is equivalent to 4 mg of docetaxel.

Example 2a: Docetaxel Nanodispersion Formulations Using Lactic Acid Buffer

Docetaxel formulations were prepared according to the general procedure outlined in Example 1a using 1 mg of lactic acid buffer (0.80 mg Lactic acid and 0.20 mg 90% sodium lactate) rather than lactic acid. These formulations are summarized Table 2a, below.

TABLE 2a Oil MCT Surfactant Solvent Docetaxel (NEOBEE ® Polysorbate 80 Polysorbate 80 PEG 300 PEG 400 No. Anhydrous Trihydrate #1053) (refined grade) (TO-10MV) (MG-300) *** (MG-400) ****  7* 4 4 95 70  8* 4 4 95 70  9A^(#) 4 4 95 70  9B^(#) 4 4 95 70  9C** 4 4 95 70  9D**^(#) 4 4 95 70 10A^(†) 4 4 95 70 10B^(†) 4 4 95 70 10C^(†) 4 4 95 70 11A^(†) 4 4 95 70 11B^(†) 4 4 95 70 11C^(†) 4 4 95 70 12A^(†) 4 4 95 70 12B^(†) 4 4 95 70 12C^(†) 4 4 95 70 13^(†) 4 4 95 70 *Dehydrated 90% sodium lactate by infrared radiation **Steam treatment (95° C. × 30 min) was applied *** Average molecule weight = 300 **** Average molecule weight = 400 ^(†)0.80 mg Lactic acid and 0.20 mg 70% sodium lactate ^(#)0.80 mg Lactic acid and 0.20 mg 90% sodium lactate

Example 3a: Stability Study

The docetaxel recovery rate, percent degradation peak (as measured by HPLC) and formulation pH were measured at one month, three months and six months at 40° C. pH was measured by mixing 3 ml of water and 750 ml of docetaxel nanodispersion sample. The results from this stability study are shown in Table 3a, below. Each formulation having lactic acid rather than either lactic acid buffer or lactic acid buffer from dehydrated sodium lactate showed a significant increase in stability.

TABLE 3a 1 month 3 months 6 months Recovery Recovery Recovery 0 month rate % rate % rate % Docetaxel Degradation (against 0 Degradation (against 0 Degradation (against 0 Degradation No. (mg) Peak % pH month) Peak % pH month) Peak % pH month) Peak % pH  6 3.722 None 3.12 98.3 None 3.10  1A 3.570 None 3.06 100.8 None 3.04 98.0 0.20 3.03 99.3 0.8 3.01  1B 3.550 None 3.09 102.1 None 3.03 98.1 0.30 3.05 100.7 0.8 3.01 (3.57*)  1C 3.570 None 3.08 101.6 None 3.03 97.9 0.30 3.04 100.5 0.8 3.02 (3.54*)  7 3.750 None 3.27 97.3 1.1 3.41 94.7 2.40 3.47 91.1 3.9 3.54  8 3.542 0.20 3.32 96.5 1.9 3.45 92.7 2.20 3.49 94.5 1.9 3.55  2A 3.800 None 3.04 97.4 None 3.04 97.3 0.40 3.03 96.9 1.4 3.07 (3.63*) (3.55*)  2B 3.784 None 3.05 99.4 None 3.03 97.6 0.30 3.06 97.0 1.0 3.10 (3.59*) (3.56*)  2C 3.810 None 3.04 98.1 None 3.03 98.1 0.40 3.02 98.0 1.1 3.07 (3.57*) (3.55*)  3A 3.805 None 3.04 97.5 None 3.05 96.8 None 3.07  3B 3.635 None 3.08 97.4 None 3.05 96.8 None 3.07  9A 3.840 None 3.44 95.3 2.1 3.57 90.2 2.50 3.60  9B 3.640 None 3.38 96.5 1.9 3.43 91.8 1.90 3.46  9C 3.670 None 3.44 95.5 1.6 3.56 88.4 3.40 3.57  9D 3.650 None 3.38 96.2 1.4 3.41 87.6 4.70 3.44 10A 3.766 0.20 96.3 2.4 3.54 90.2 2.90 3.53 88.7 2.1 3.60 (3.77*) 10B 3.722 0.20 96.7 2.1 3.57 90.9 2.80 3.57 87.4 2.2 3.63 (3.78*) 10C 3.750 0.20 96.4 2.5 3.56 91.3 2.80 3.56 88.2 2.2 3.62 (3.76*) 11A 3.740 0.20 96.9 2.0 3.52 91.2 3.40 3.55 86.4 2.9 3.61 (3.77*) 11B 3.654 0.20 97.9 1.9 3.55 91.6 3.10 3.55 88.0 2.7 3.61 (3.78*) 11C 3.732 0.20 97.0 2.2 3.55 90.7 3.30 3.55 88.2 2.6 3.62 (3.80*) 12A 3.615 0.10 3.64* 95.4 2.8 3.39 95.6 4.00 3.39 87.5 3.5 3.50 (3.70) 12B 3.627 0.20 3.62* 95.6 2.8 3.38 96.4 4.00 3.37 89.3 3.3 3.50 (3.69*) 12C 3.078 15.00  3.61* 95.6 18.4  3.38 96.5 20.30 3.37 87.2 20.3 3.50 (3.69*) 13 3.660 0.10 94.7 2.6 3.41 93.8 4.00 3.42 86.7 3.6 3.50 (3.70*)  4A 3.676 None 3.03 100.4 None 3.03 101.0 0.1 3.07 (3.54*)  4B 3.618 0.10 3.02 101.4 None 3.02 101.4 0.1 3.05 (3.56*)  4C 3.632 0.10 3.01 100.4 None 3.01 102.4 0.1 3.11 (3.57*)  5A 3.746 None 3.04 101.9 None 3.04 102.2 0.1 3.06 (3.57*)  5B 3.770 None 3.04 101.7 None 3.02 102.3 0.1 3.05 (3.54*)  5C 3.732 None 3.02 102.3 None 3.01 101.6 0.1 3.02 (3.54*) *pH of docetaxel diluted solution.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. 

The invention claimed is:
 1. A method of treating a cellular proliferative disease, comprising administering to a subject in need thereof a non-aqueous, ethanol-free docetaxel liquid formulation comprising: docetaxel in an amount ranging from about 0.5 to about 5 wt %; an oil in an amount ranging from about 1 to about 5 wt %; a non-ionic surfactant; a non-aqueous solvent, which is polyethylene glycol having an average molecular weight of less than about 1000, in an amount ranging from about 20 to about 60 wt %; and an organic acid component; wherein the organic acid component is soluble in the non-aqueous solvent and the amount by weight of non-ionic surfactant is equal to or greater than the amount by weight of non-aqueous solvent.
 2. The method according to claim 1, wherein the docetaxel is present in about 2 wt %.
 3. The method according to claim 2, wherein the docetaxel is anhydrous docetaxel.
 4. The method according to claim 2, wherein the docetaxel is docetaxel trihydrate.
 5. The method according to claim 1, wherein the oil is selected from the group consisting of synthetic oils, vegetable oils, tocopherols and combinations thereof.
 6. The method according to claim 5, wherein the oil is selected from the group consisting of soybean oil, olive oil, sesame oil, corn oil, a medium chain triglyceride, a tocopherol or derivative thereof and combinations thereof.
 7. The method according to claim 6, wherein the oil is soybean oil.
 8. The method according to claim 1, wherein the non-ionic surfactant is present in an amount ranging from about 40 to about 75 wt %.
 9. The method according to claim 8, wherein the non-ionic surfactant is present in an amount ranging from about 50 to about 60 wt %.
 10. The method according to claim 1, wherein the non-ionic surfactant is polysorbate
 80. 11. The method according to claim 1, wherein the non-aqueous solvent is present in an amount ranging from about 35 to about 45 wt %.
 12. The method according to claim 1, wherein the organic acid component is present in an amount ranging from 0.5 to 1 wt %.
 13. The method according to claim 12, wherein the organic acid component is an organic acid selected from the group consisting of lactic acid and acetic acid.
 14. The method according to claim 13, wherein the organic acid is lactic acid.
 15. The method according to claim 1, wherein the amount by weight of oil is equal to or less than about 2.5 times the amount of taxane.
 16. The method according to claim 1, wherein in the non-aqueous, ethanol-free docetaxel liquid formulation: the docetaxel is docetaxel trihydrate; the oil is soybean oil the non-ionic surfactant is polysorbate 80 in an amount ranging from about 50 to about 60 wt %; the non-aqueous solvent is polyethylene glycol in an amount ranging from about 35 to about 45 wt %; and the organic acid component is present in an amount ranging from 0.3 to 1 wt %.
 17. The method according to claim 1, wherein the ethanol-free docetaxel liquid formulation has a pH of less than about 3.5.
 18. The method according to claim 1, wherein the formulation is stable for more than 6 months at 40° C.
 19. The method according to claim 1, wherein the formulation exhibits a docetaxel recovery rate of greater than 95% after 6 months at 40° C.
 20. The method according to claim 1, wherein the formulation forms particles of less than about 20 nm when combined with an aqueous medium.
 21. The method according to claim 1, wherein the ethanol-free docetaxel liquid formulation is administered to the subject by: (a) combining the ethanol-free docetaxel formulation with an aqueous medium to provide an ethanol-free docetaxel diluted solution; and (b) administering the ethanol-free docetaxel diluted solution to the subject. 